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Determination of methylation in the promotor regions of genes, that control metabolism of 5-FU
Bendová, Petra ; Vodička, Pavel (advisor) ; Václavíková, Radka (referee)
Several malignant diseases, such as colorectal, pancreatic, breast or ovarial cancers, are primarily treated with cytostatics 5-fluorouracil (5-FU). 5-FU undergoes biotransformation in human body and arising metabolites induce the damage and subsequent apoptosis in the target cells. The main aim of this diploma Thesis was the determination of methylation in promoter regions of 14 candidate genes participating on 5-FU biotransformation: TK1, PPAT, RRM1, RRM2, UCK2, UCK1, UMPS, TYMP, UPP1, UPP 2 SLC29A1, UPB1, DPYS and DPYD. We hypothesize that the methylation in promoter regions regulates mRNA transcription of the above candidate genes. We have conducted appropriate analyses in 128 colorectal cancer patients, for whom both tumor and nonmalignant adjacent tissues were available. Sample processing and analysis involved DNA isolation, bisulfite conversion of unmethylated cytosines to corresponding uracils, methylation-specific analysis of melting curves with high resolution for theproper methylation analysis and gel electrophoresis to separate PCR products. For the majority of the studied genes (TK1, PPAT, RRM1, RRM2, UCK2, UCK1, UMPS, TYMP, UPP1, SLC29A1 and DPYD) we did not detect any aberrant methylation in promoter regions. In genes DPYS, UPB1 and UPP2 we recorded various degree of promoter...
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